Microtubules as platforms for probing liquid-liquid phase separation in cells - application to RNA-binding proteins.
J Cell Sci
; 131(11)2018 06 11.
Article
en En
| MEDLINE
| ID: mdl-29728455
ABSTRACT
Liquid-liquid phase separation enables compartmentalization of biomolecules in cells, notably RNA and associated proteins in the nucleus. Besides having critical functions in RNA processing, there is a major interest in deciphering the molecular mechanisms of compartmentalization orchestrated by RNA-binding proteins such as TDP-43 (also known as TARDBP) and FUS because of their link to neuron diseases. However, tools for probing compartmentalization in cells are lacking. Here, we developed a method to analyze the mixing and demixing of two different phases in a cellular context. The principle is the following RNA-binding proteins are confined on microtubules and quantitative parameters defining their spatial segregation are measured along the microtubule network. Through this approach, we found that four mRNA-binding proteins, HuR (also known as ELAVL1), G3BP1, TDP-43 and FUS form mRNA-rich liquid-like compartments on microtubules. TDP-43 is partly miscible with FUS but immiscible with either HuR or G3BP1. We also demonstrate that mRNA is essential to capture the mixing and demixing behavior of mRNA-binding proteins in cells. Taken together, we show that microtubules can be used as platforms to understand the mechanisms underlying liquid-liquid phase separation and their deregulation in human diseases.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Células
/
Proteínas de Unión al ARN
/
Microscopía Fluorescente
/
Microtúbulos
Tipo de estudio:
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
J Cell Sci
Año:
2018
Tipo del documento:
Article
País de afiliación:
Francia